Crystalline polymorphic forms of an antidiabetic compound

ABSTRACT

The present invention relates to polymorphic forms of a compound of formula A: 
                         
This compound is useful as a glucagon receptor antagonist and serves as a pharmaceutically active ingredient for the treatment of type 2 diabetes and related conditions, such as hyperglycemia, obesity, dyslipidemia, and the metabolic syndrome. Hydrates, hemihydrates, anhydrates and similar polymorphic forms are included.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a continuation application of U.S.application Ser. No. 14/186,829, filed Feb. 21, 2014, which is adivisional application of U.S. application Ser. No. 13/144,031 (now U.S.Pat. No. 8,697,740), filed Sep. 23, 2011 as a U.S. National Phaseapplication under 35 U.S.C. §371 of PCT/US2010/020460, filed Jan. 8,2010, which claims priority from U.S. provisional Application Ser. No.61/204,886, filed Jan. 12, 2009. The present application claims priorityto the foregoing applications.

FIELD OF THE INVENTION

The present invention relates to novel polymorphic forms of a compoundof formula A. This compound is useful as a pharmaceutically activeingredient for the treatment of type 2 diabetes and related conditions,such as hyperglycemia, obesity, dyslipidemia, and the metabolicsyndrome.

BACKGROUND OF THE INVENTION

Type 2 diabetes remains a serious medical problem. There is an ongoingneed for new treatments that are more effective and that have fewer sideeffects. Glucagon receptor antagonists are important upcomingmedications for the treatment of type 2 diabetes and the presentcompound is particularly useful in this regard.

SUMMARY OF THE INVENTION

The present invention relates to polymorphic forms of a compound offormula A:

The compound is also known asN-(4-{(1S)-1-[(R)-(4-chlorophenyl)(7-fluoro-5-methyl-1H-indol-3-yl)methyl]butyl}benzoyl)-β-alanine.Compound A has been disclosed in a published PCT patent application,WO2008/042223 published on Apr. 10, 2008. Polymorphic forms of thecompound that are particularly useful in the preparation ofpharmaceutical products are described herein.

The invention also relates to pharmaceutical compositions comprising thepolymorphic forms described herein, methods for the preparation thereofand the like

BRIEF DESCRIPTION OF THE DRAWINGS

The invention is described in connection with the drawings appendedhereto, in which:

FIG. 1 is the X-ray powder diffraction (XRPD) pattern for the free acidhydrate polymorphic Form I of Compound A;

FIG. 2 is the solid-state 19 F CPMAS NMR spectrum of the free acidhydrate polymorphic Form I of Compound A;

FIG. 3 is the 13C solid state NMR of the free acid hydrate polymorphicForm I of Compound A;

FIG. 4 is the X-ray powder diffraction (XRPD) pattern of anhydrous freeacid polymorphic Form I of Compound A;

FIG. 5 is the 19F solid state NMR of anhydrous free acid polymorphicForm I of Compound A;

FIG. 6 is the X-ray diffraction pattern of the crystalline anhydrateForm II of Compound A;

FIG. 7 is the Thermogravimetric analysis curve of the crystallineanhydrate From II of Compound A;

FIG. 8 is the Differential Scanning calorimetry curve of the crystallineanhydrate Form II of Compound A;

FIG. 9 is the Solid State C-13 CPMAS NMR spectrum for the crystallineanhydrate Form II of Compound A;

FIG. 10 is the Fluorine-19 Single Pulse Excitation MAS spectrum ofAnhydrate II of Compound A.

FIG. 11 is the X-ray Powder Diffraction pattern of the crystallineanhydrate Form III of Compound A;

FIG. 12 is the Thermogravimetric analysis curve of the crystallineanhydrate Form III of Compound A;

FIG. 13 is the Differential Scanning calorimetry curve of thecrystalline anhydrate Form III of Compound A;

FIG. 14 is the Solid State C-13 CPMAS NMR spectrum for the crystallineanhydrate Form III of Compound A;

FIG. 15 is the Fluorine-19 Single Pulse Excitation MAS spectrum ofcrystalline anhydrate Form III of Compound A, and

FIG. 16 is an X Ray Powder Diffraction pattern for comparison purposesof Compound A containing a mixture of amorphous product and polymorphs.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a process for the preparation ofcrystallineN-(4-{(1S)-1-[(R)-(4-chlorophenyl)(7-fluoro-5-methyl-1H-indol-3-yl)methyl]butyl}benzoyl)-β-alanineof structural formula A:

and specifically polymorphic forms thereof, including solvates.

Crystal forms are convenient for the preparation and isolation ofcompound A with an upgrade in purity, and represent a convenientscalable way to produce high purity compound A. Crystalline forms wereidentified including the crystalline free base of Compound A as analcohol solvate and various crystalline salt forms of Compound A areincluded as well. These crystalline salts of compound A are novel andhave improved physiochemical properties, such as purity, stability andease of formulation that render them particularly suitable for themanufacture of pharmaceutical dosage forms. Discovery of crystallineforms allowed for the facile purification, isolation, and formulation ofcompound A.

The abbreviations in the table below have the following meanings:

Me = methyl Et = ethyl t-Bu = t-butyl IPA = isopropyl alcohol Ac =Acetyl THF = tetrahydrofuran DCM = dichloromethane DIEA =diisopropylethylamine DMF = dimethylformamide DPE phos = Bis(2-diphenylphosphinophenyl)ether IPAC = isopropylacetate MTBE = methylt-butyl ether BINAP = 2,2′-bis(diphenylphosphino)- Tol-BINAP =2,2′-bis(di-p- 1,1′-binaphthyl tolylphosphino)-1,1′-binaphthyl

One aspect of the invention that is of interest relates to process forsynthesizing a compound of formula A:

comprising deprotecting a compound of formula 11a:

wherein P¹ and P² represent protecting groups, to produce a compound offormula A.

Another aspect of the invention relates to a process for synthesizing acompound of formula A:

comprising reacting a compound of formula 10a:

wherein P¹ represents a protecting group with a beta alanine ester ofthe formulaH₂NCH₂CH₂CO₂P²wherein P² represents a protecting group, with a peptide coupling agent,to produce a compound of formula 11a:

and deprotecting compound 11a to produce a compound of formula A.

Another aspect of the invention relates to a A process for the synthesisof a compound of formula A:

comprising reacting a compound of the formula 11:

with base to produce a compound of formula A.

Another aspect of the invention relates to a process as described abovewherein the base is NaOH.

Another aspect of the invention relates to a process for synthesizing acompound of formula A as shown above.

A compound of formula 10a is reacted with N,N-carbonyldiimidazole (CDI)or another peptide coupling reagent and a beta-alanine esterH₂NCH₂CH₂CO₂P² to produce a compound of formula 11a. P¹ represents aprotecting group suitable for protection of the indole nitrogen atom.Representative examples include nosyl, benzyl, tolyl and similar groups,with nosyl being preferred. Suitable protecting groups for the betaalanine ester, P², include small alkyl groups, such as methyl.Thereafter deprotection is undertaken to remove P¹ and P², thus forminga compound of formula A.

A synthetic route for the compound of formula A has been disclosed inWO2008/042223 published on Apr. 10, 2008, hereby incorporated byreference, and is set forth below. In the present invention, seedcrystals of compound were generated from lab scale runs and formedbefore or after chromatographic purification.

Intermediate 1 Racemic4-[2-(4-chlorophenyl)-1-propylpent-4-en-1-yl]benzoic acid

Step A. tert-Butyl 4-[2-(4-chlorophenyl)-2-oxoethyl]benzoate

A THF solution (200 ml) containing t-butyl 4-bromobenzoate (19.9 g, 77.6mmol), 4-chloroacetophenone (10 g, 64.7 mmol), Pd₂dba₃ (1.19 g, 1.29mmol), BINAP (1.6 g, 2.58 mmol) and NaOtBu (8.7 g, 90.6 mmol) wasrefluxed under an argon atmosphere for approximately 5 hours. Thesolution was concentrated and then partitioned between EtOAc and water.The organic phase was washed with water, brine and dried over Na₂SO₄.The filtered solution was concentrated and the residue purified bysilica gel chromatography using a hexanes/EtOAc gradient to give thetitle compound. ¹H NMR (500 MHz, CDCl₃): δ 7.95 (d, J=8.5 Hz, 2H); 7.93(d, J=8.7 Hz, 2H); 7.43 (d, J=8.3 Hz, 2H); 7.29 (d, J=8.2 Hz, 2H); 4.30(s, 2H); 1.58 (s, 9H). LC1 4.01 min. (M-tBu+H)=275

Step B. tert-Butyl 4-[1-(4-chlorobenzoyl)butyl]benzoate

KOtBu (2.55 g, 22.7 mmol) was added to a cooled (ice bath) THF solution(40 ml) containing the intermediate from Step A (5.0 g, 15.15 mmol).After 10 minutes n-propyl iodide (3 ml, 30.3 mmol) was added dropwise.The ice bath was removed and the reaction was monitored by MS-HPLCanalysis. The solution was then partitioned (<1 hour) between EtOAc andwater. The organic phase was washed with water, brine and dried overNa₂SO₄. The filtered solution was concentrated and the residue purifiedby silica gel chromatography using a hexanes/EtOAc gradient to give thetitle compound. ¹H NMR (400 MHz, CDCl₃): δ 7.90 (d, J=7.8 Hz, 2H); 7.84(d, J=8.6 Hz, 2H); 7.33 (d, J=8.6 Hz, 2H); 7.31 (d, J=8.3 Hz, 2H); 4.51(t, J=7.2 Hz, 1H); 2.18-2.08 (m, 1H); 1.84-1.68 (m, 1H); 1.54 (s, 9H);1.38-1.18 (m, 2H); 0.90 (t, J=7.3 Hz, 3H). LC1 4.43 min. (M-tBu+H)=317

Step C. tert-Butyl 4-{1-[(4-chlorophenyl)(hydroxy)methyl]butyl}benzoate

NaBH₄ (0.5 g, 13.21 mmol) was added in portions to a MeOH solution (40ml) containing the intermediate from Step B (3.78 g, 10.16 mmol). Afterstirring for 1 hour the solution was concentrated and the residuepartitioned between EtOAc and water. The organic phase was washed withwater, brine and dried over Na₂SO₄. The filtered solution wasconcentrated and the residue purified by silica gel chromatography usinga hexanes/EtOAc gradient to give the title compound as a >10:1 ratio ofdiastereomers. ¹H NMR (400 MHz, CDCl₃): δ 7.93 (d, J=8.3 Hz, 2H); 7.28(d, J=8.4 Hz, 2H); 7.23 (d, J=8.4 Hz, 2H); 7.18 (d, J=8.4 Hz, 2H); 4.73(d, J=7.8 Hz, 1H); 2.89-2.83 (m, 1H); 1.58 (s, 9H); 1.57-1.56 (m, 1H);1.41-1.33 (m, 1H); 1.09-0.91 (m, 2H); 0.72 (t, J=7.3 Hz, 3H). LC1 4.22min. (M-tBu-OH+H)=301

Step D. 4-[2-(4-Chlorophenyl)-1-propylpent-4-en-1-yl]benzoic acid

A 1,2-dichloroethane (DCE) (20 ml) solution containing the intermediatefrom Step C (1.81 g, 4.84 mmol), allyl trimethylsilane (6.2 ml, 38.7mmol) and boron trifluoride etherate (1.84 ml, 14.5 mmol) was heated at80° C. for 1.5 hours. The solution was cooled to room temperature andmethanol (10 ml) was slowly added. The solution was then concentratedand the residue partitioned between EtOAc and aqueous 1N HCl. Theorganic phase was washed with water, brine and dried over Na₂SO₄. Thefiltered solution was concentrated to give the title compound (as a ca3:1 mixture of diastereomers) which was used without furtherpurification. A portion was purified for spectral analysis. Data is forthe major diastereomer ¹H NMR (400 MHz, CDCl₃): δ 8.07 (d, J=8.3 Hz,2H); 7.30 (d, J=5.7 Hz, 2H); 7.28 (d, J=5.4 Hz, 2H); 7.08 (d, J=8.3 Hz,2H); 5.42-5.32 (m, 1H); 4.79-4.66 (m, 2H); 2.83-2.77 (m, 2H); 2.11-2.05(m, 2H); 1.43-1.29 (m, 2H); 1.00-0.80 (m, 2H); 0.68 (t, J=7.3 Hz, 3H).LC1 4.08 min.

(M+H)=343

NMR experiments (NOE) on advanced compounds (see EXAMPLE 1) derived fromINTERMEDIATE 1 established the relative stereochemistry of the minor andmajor diastereomers of INTERMEDIATE 1 as:

Intermediate 24-[(1S,2R)-2-(4-chlorophenyl)-1-propylpent-4-en-1-yl]benzoic acid

Step A.2-(4-Bromophenyl)-N-[(1R,2R)-2-hydroxy-1-methyl-2-phenylethyl]-N-methylacetamide

Pivaloyl chloride (7.8 ml, 63.3 mmol) was added dropwise to a DCM/THFsolution (100 ml/20 ml) containing 4-bromophenylacetic acid (13.59 g,63.2 mmol). DIEA (11.0 ml, 63.1 mmol) was then added dropwise(exotherm). After stirring at room temperature for 1 hour the solutionwas poured slowly into a DCM/THF solution (100 ml/20 ml) containing(1R,2R)-(−)-pseudoephedrine (10.5 g, 63.5 mmol) and DIEA (11.0 ml, 63.1mmol). After stirring overnight at room temperature the solution wasconcentrated and the residue partitioned between EtOAc and water. Theorganic phase was washed with aqueous 1N NaOH (2×), aqueous 1N HCl (3×),brine and dried over MgSO₄. The solution was filtered and concentrated.The oil residue was diluted with 100 ml of toluene and concentrated. Theresidue was then dissolved in ethyl ether and triturated with hexanes togive the title compound as a white solid. The compound is a 3:1 mixtureof amide rotational isomers by proton NMR: ¹H NMR (400 MHz, asteriskdenotes minor rotamer, CDCl₃): δ 7.42 (d, J=8.3 Hz, 2H); 7.39-7.27 (m,5H); 7.11*(d, J=8.4 Hz, 2H); 7.04 (d, J=8.3 Hz, 2H); 4.64-4.42 (m, 1H);4.07-3.94 (m, 1H); 3.82-3.70 (m, 1H); 2.94*(s, 3H); 3.63 (s, 2H); 2.82(s, 3H); 1.12 (d, J=7.0 Hz, 3H); 0.86*(d, 3H, J=7.0 Hz). LC1 3.23 min.(M+H)=362

Step B.2-(4-Bromophenyl)-N-[(1R,2R)-2-hydroxy-1-methyl-2-phenylethyl]-N-methylpentanamide

THF (40 ml) was added to dry lithium chloride (8 g, 189 mmol) anddiisopropyl amine (9.2 ml, 65.6 mmol) under an argon atmosphere. Thesuspension was cooled to −78° C. and n-BuLi (1.6M in hexanes, 37.9 ml,60.6 mmol) was added dropwise. After stirring for 5 minutes the solutionwas warmed to 0° C. After 5 minutes the solution was cooled to −78° C.and a THF solution (45 ml) containing the intermediate from Step A(10.56 g, 29.15 mmol) was added dropwise. The solution was then stirredat −78° C. for 1 hour and then warmed to 0° C. After 15 minutes n-propyliodide (4.3 ml, 44.1 mmol) was added dropwise. The solution was stirredat 0° C. for approximately 2 hours. To the reaction mixture was addedsaturated aqueous NH₄Cl and EtOAc. The phases were separated and theaqueous phase extracted with EtOAc. The combined organic phases weredried over Na₂SO₄, filtered and concentrated. The oil residue wasdissolved in ethyl ether/hexanes (4/6) and filtered through a short plugof silica gel. The filtered solution was concentrated to give the titlecompound. The compound is a 3:1 mixture of amide rotational isomers byproton NMR: ¹H NMR (400 MHz, asterisk denotes minor rotamer, CDCl₃): δ7.42 (d, J=8.4 Hz, 2H); 7.41-7.27 (m, 5H); 7.08 (d, J=8.4 Hz, 2H); 4.56(q, J=6.7 Hz, 1H); 4.42 (br s 1H); 4.17-4.01*(m, 1H); 3.85*(t, J=7.1 Hz,1H); 3.55 (t, J=7.2 Hz, 1H); 3.00* (s, 3H); 2.72 (s, 3H); 2.07-1.92 (m,1H); 1.69-1.58 (m, 1H); 1.33-1.13 (m, 2H); 1.11 (d, J=7.0 Hz, 3H); 0.88(t, J7.3 Hz, 3H): 0.58* (d, J=6.9 Hz, 3H). LC1 3.76 min. (M+H)=404

Step C. 2-(4-Bromophenyl)-1-(4-chlorophenyl)pentan-1-one

n-Butyl lithium (1.0M in THF, 59 ml, 94.5 mmol) was added dropwise to a−78° C. THF solution (200 ml) containing 4-chloro bromobenzene (22.63 g,118.2 mmol) under an argon atmosphere. After 10 minutes a THF solution(30 ml) of the intermediate from Step B (15.88 g, 39.4 mmol) was addeddropwise. The solution was warmed to 0° C. and stirred for 30 minutes.Diisopropylamine (5.6 ml, 39.4 mmol) was then added dropwise. After 10minutes the reaction solution was diluted with 200 ml of AcOH/ethylether (1/10 by volume). The mixture was partitioned between EtOAc andsaturated aqueous NaHCO₃ (foaming). The organic phase was washed withsaturated aqueous NaHCO₃, water, brine and dried over Na₂SO₄. Thefiltered solution was concentrated and the residue purified by silicagel chromatography using hexanes/EtOAc gradient to give the titlecompound. ¹H NMR (500 MHz, CDCl₃): δ 7.86 (d, 2H, J=8.5 Hz); 7.41 (d,2H, J=8.5 Hz); 7.37 (d, 2H, J=8.5 Hz); 7.15 (d, 2H, J=8.5 Hz); 4.45 (t,J=7.3 Hz, 1H); 2.15-2.07 (m, 1H); 1.81-1.73 (m, 1H); 1.33-1.19 (m, 2H);0.91 (t, J=7.4 Hz, 3H). LC1 4.25 min. Not ionized

Step D. 2-(4-Bromophenyl)-1-(4-chlorophenyl)pentan-1-ol

Sodium borohydride (917 mg, 24.25 mmol) was added to a MeOH solution (25ml) containing the intermediate from Step C (6.53 g, 18.66 mol). Afterstirring for 1 hour at room temperature the solution was concentratedand the residue partitioned between water and EtOAc. The organic phasewas washed with water, brine and dried over Na₂SO₄. The filteredsolution was concentrated to give the title compound as an 8:1 mixtureof diastereomers which was used in the next step without furtherpurification. ¹H NMR for major diastereomer (500 MHz, CDCl₃): δ 7.44 (d,J=8.1 Hz, 2H); 7.30 (d, J=8.5 Hz, 2H); 7.19 (d, J=8.5 Hz, 2H); 7.07 (d,J=8.1 Hz, 2H); 4.71-4.68 (m, 1H); 2.81-2.74 (m, 1H); 1.56-1.48 (m, 1H);1.42-1.32 (m, 1H); 1.12-0.95 (m, 2H); 0.75 (t, J=7.3 Hz, 3H). LC1 4.00min. (M-OH)=335

Step E. 1-Bromo-4-[2-(4-chlorophenyl)-1-propylpent-4-en-1-yl]benzene

The title compound was prepared from the intermediate from Step D usingthe conditions described in Step D. The title compound is obtained as a2.1:1 mixture of diastereomers. ¹H NMR for major diastereomer (500 MHz,CDCl₃): δ 7.44 (d, J=8.5 Hz, 2H); 7.28 (d, J=8.3 Hz, 2H); 7.05 (d, J=8.2Hz, 2H); 7.02 (d, J=8.4 Hz, 2H); 5.46-5.35 (m, 1H); 4.82-4.71 (m, 2H);2.77-2.62 (m, 2H); 2.14-2.02 (m, 2H); 1.35-1.25 (m, 2H); 1.05-0.89 (m,2H); 0.67 (t, J=7.3 Hz, 3H). LC1 4.66 min. Not ionized

Step F. n-Butyl 4-[2-(4-chlorophenyl)-1-propylpent-4-en-1-yl]benzoate

An n-butanol solution (5 ml) containing the intermediate from Step E(108 mg, 0.286 mmol), DIEA (0.15 ml, 0.86 mmol) and PdCl₂(PPh₃)₂ (376mg, 0.06 mmol) was heated at 115° C. under a carbon monoxide atmosphere(balloon). After 1 hour the solution was cooled and concentrated. Theresidue was dissolved in EtOAc and filtered. The residue was usedwithout purification in the next step. A portion was purified forspectral analysis. ¹H NMR for major diastereomer (500 MHz, CDCl₃): δ8.00 (d, J=8.3 Hz, 2H); 7.28 (d, J=8.4 Hz, 2H); 7.23 (d, J 8.3 Hz, 2H):7.07 (d J=8.4 Hz, 2H); 5.42-5.31 (m, 1H); 4.77-4.66 (m, 2H); 4.33 (t,J=6.6 Hz, 2H); 2.80-2.75 (m, 2H); 2.10-2.06 (m, 2H); 1.81-1.68 (m, 2H);1.41-1.24 (m, 4H); 0.99 (t, J=7.4 Hz, 3H); 0.98-0.86 (m, 4H); 0.67 (t,J=7.3 Hz, 3H). LC1 4.73 min. (M+H)=399

Step G. 4-[(1S,2R)-2-(4-chlorophenyl)-1-propylpent-4-en-1-yl]benzoicacid

A THF/MeOH/water (8 ml/8 ml/3 ml) solution containing the intermediatefrom Step F (790 mg, 1.98 mmol) and lithium hydroxide monohydrate (406mg, 9.90 mmol) was stirred overnight at room temperature. The solutionwas concentrated and the nonvolatile portion was partitioned betweenaqueous 2N hydrochloric acid and EtOAc. The organic phase was dried overNa₂SO₄, filtered and concentrated to give the title compound. ¹H NMR(500 MHz, DMSO-d₆): δ 7.90 (d, J=8.2 Hz, 2H); 7.39 (d, J=8.5 Hz, 2H);7.36 (d, J=8.5 Hz, 2H); 7.26 (d, J=8.4 Hz, 2H); 5.36-5.26 (m, 1H);4.71-4.60 (m, 2H); 2.94-2.84 (m, 2H); 2.13-2.07 (m, 1H); 1.95-1.87 (m,1H); 1.42-1.34 (m, 1H); 1.19-1.11 (m, 1H); 0.85-0.77 (m, 2H); 0.60 (t,J=7.3 Hz, 3H). LC3 2.57 min (M+H) 343

Alternatively, the title compound can be prepared from the intermediatefrom Step E. A pentane solution of t-BuLi (1.7M, 3.08 ml, 5.23 mmol) wasadded dropwise to a THF solution (20.1 ml) of the intermediate from StepE (760 mg, 2.01 mmol) cooled to −78° C. After 5 minutes, CO₂ gas wasbubbled for a half minute through the solution. The cooling bath wasremoved and the solution was warmed to room temperature. The solutionwas then diluted with aqueous 2N HCl and extracted with EtOAc (2×). Thecombined organic phases were dried over Na₂SO₄, filtered andconcentrated to give the title compound.

The absolute stereochemistry of the minor and major diastereomers ofINTERMEDIATE 2 is shown below. This assignment is based on the knownconfiguration of the n-propyl substituted carbon, which is derived fromthe (−)-pseudoephedrine, and NMR experiments (NOE). See WO2008/042223.

Intermediate 3 MethylN-{4-[2-(4-chlorophenyl)-1-propylpent-4-en-1-yl]benzoyl}-β-alaninate

A DMF solution (20 ml) containing INTERMEDIATE 1 (1.66 g, 4.84 mmol),methyl β-alaninate hydrochloride (1.01 g, 7.26 mmol), DIEA (4.3 ml, 24.2mmol) and BOP (3.21 g, 7.26 mmol) was stirred at room temperature for1.5 hours. The solution was diluted with EtOAc and washed with water,brine and dried over Na₂SO₄. The filtered solution was concentrated andthe residue purified by silica gel chromatography using a hexanes/EtOAcgradient to give the title compound. ¹H NMR for the major diastereomer(500 MHz, CDCl₃): δ 7.72 (d, J=8.2 Hz, 2H); 7.28 (d, J=8.3 Hz, 2H); 7.22(d, J=8.2 Hz); 7.07 (d, J=8.4 Hz, 2H); 6.85-6.81 (m, 1H); 5.41-5.31 (m,1H); 4.77-4.66 (m, 2H); 3.75-3.70 (m, 2H); 3.73 (s, 3H); 2.81-2.72 (m,2H); 2.67 (t, J=5.9 Hz, 2H); 2.10-2.05 (m, 2H); 1.40-1.29 (m, 2H);0.98-0.85 (m, 2H); 0.66 (t, J=7.3 Hz, 3H). LC1 4.03 min. (M+H)=428

Intermediate 4 MethylN-{4-[2-(4-chlorophenyl)-4-oxo-1-propylbutyl]benzoyl}-β-alaninate

Ozone was purged through a chilled (−78° C.) DCM solution (20 ml)containing INTERMEDIATE 3 (1.59 g, 3.72 mmol). The ozone purge wasmaintained until an excess of ozone was observed (blue color, <10minutes). The solution was then purged with nitrogen to dissipate theexcess ozone. To the solution was added dimethylsulfide (1 ml) followedby triphenylphosphine (977 mg, 3.72 mmol). The solution was warmed toroom temperature and stirred for approximately 2 hours. The solution wasconcentrated and the residue purified by silica gel chromatography usinga hexanes/EtOAc gradient to give the title compound. ¹H NMR for themajor diastereomer (500 MHz, CDCl₃): δ 9.34 (s, 1H); 7.73 (d, J=8.2 Hz,2H); 7.30 (d, J=8.3 Hz, 2H); 7.23 (d, J=8.0 Hz, 2H); 7.16 (d, J=8.4 Hz,2H); 6.87-6.83 (broad s, 1H); 3.72 (s, 3H); 3.75-3.71 (m, 2H); 3.36-3.31(m, 1H); 2.80-2.72 (m, 1H); 2.69-2.63 (m, 2H); 2.61-2.52 (m, 1H); 2.38(dd, J=3.9, 17.1 Hz, 1H); 1.45-1.28 (m, 2H); 1.06-0.78 (m, 2H); 0.66 (t,J=7.3 Hz, 3H). LC1 3.55 min. (M+H)=430

Example 1N-(4-{(1S)-1-[(R)-(4-chlorophenyl)(7-fluoro-5-methyl-1H-indol-3-yl)methyl]butyl}benzoyl)-β-alanine

Step A. MethylN-(4-{(1S)-1-[(R)-(4-chlorophenyl)(7-fluoro-5-methyl-1H-indol-3-yl)methyl]butyl}benzoyl)-β-alaninate(Compound A)

An acetic acid solution (10 ml) of methylN-{4-[2-(4-chlorophenyl)-4-oxo-1-propylbutyl]benzoyl}-β-alaninate,INTERMEDIATE 4, (757 mg, 1.76 mmol), ZnCl₂ (3.1M in AcOH, 1.7 ml, 5.27mol) and 2-fluoro-4-methylphenylhydrazine hydrochloride (374 mg, 2.1mmol) was heated at 80° C. for 45 minutes. The solution was concentratedand the residue partitioned between EtOAc and water. The organic phasewas washed with water (2×), brine (2×) and dried over Na₂SO₄. Thesolution was filtered, concentrated and the residue purified by silicagel chromatography using a hexanes/ethyl acetate gradient to give thetitle compound. Data for the major diastereomer: ¹H NMR (500 MHz,CD3CN): δ 9.11 (s, 1H); 7.54 (d, J=8.2 Hz, 2H); 7.48 (d, J=8.5 Hz, 2H);7.38 (d, J=8.2 Hz, 2H); 7.30 (d, J=8.4 Hz, 2H); 7.15 (d, J=2.5 Hz, 1H);7.11 (s, 1H); 7.02-6.97 (m, 1H); 6.59 (d, J=12.3 Hz, 1H); 4.49 (d,J=11.6 Hz, 1H); 3.60 (s, 3H); 3.56-3.48 (m, 3H); 2.52 (t, J=6.8 Hz, 2H);2.32 (s, 3H); 1.49-1.35 (m, 2H); 1.04-0.90 (m, 2H); 0.69 (t, J=7.4 Hz,3H). LC1=3.94 min. (M+H)=535. Chiral LC1 (1% to 15% EtOH/heptane over 25min, 15% EtOH/heptane isocratic >25 min) retention time=28.38 minutes.The material also contains ca 2% by area of the enantiomer. Chiral LC1(1% to 15% EtOH/heptane over 25 min, 15% EtOH/heptane isocratic >25 min)retention time=26.88 minutes.

Step B.N-(4-{(1S)-1-[(R)-(4-Chlorophenyl)(7-fluoro-5-methyl-1H-indol-3-yl)methyl]butyl}benzoyl)-β-alanine

The isomers obtained in Step A were hydrolyzed using the conditionsdescribed in INTERMEDIATE 2, Step G. The crude hydrolysis was purifiedby HPLC to give the title compounds.

Data for the Minor Diastereomer:

¹H NMR (400 MHz, CD₃CN): δ 9.39 (s, 1H); 7.56 (d, J=8.0 Hz, 2H); 7.37(d, J=2.4 Hz, 1H); 7.33 (s, 1H); 7.29 (d, J=8.0 Hz, 2H); 7.20 (d, J=8.4Hz); 7.07 (broad s, 1H); 7.01 (d, J=8.4 Hz, 2H); 6.72 (d, J=12.4 Hz,1H); 4.45 (d, J=11.6 Hz); 3.65-3.55 (m, 1H); 3.52 (q, J=6.4 Hz, 2H);2.55 (t, J=6.8 Hz, 2H); 2.40 (s, 3H); 1.84-1.73 (m, 1H); 1.63-1.52 (m,1H); 1.10-0.93 (m, 2H); 0.71 (t, J=7.2 Hz, 3H). LC1=3.66 min. (M+H)=521.

Data for the major diastereomer: ¹H NMR (500 MHz, CD₃CN): δ 9.11 (s,1H); 7.56 (d, J=8.2 Hz, 2H); 7.49 (d, J=8.4 Hz, 2H); 7.39 (d, J=8.2 Hz,2H); 7.31 (d, J=8.4 Hz, 2H); 7.16 (d, J=2.4 Hz, 1H); 7.12 (s, 1H); 7.04(s, 1H); 6.60 (d, J=12.2 Hz, 1H); 4.50 (d, J=11.6 Hz, 1H); 3.58-3.53 (m,1H); 3.50 (q, J=6.4 Hz, 2H); 2.53 (t, J=6.6 Hz, 2H); 2.33 (s, 3H);1.51-1.37 (m, 2H); 0.99-0.92 (m, 2H); 0.70 (t, J=7.3 Hz, 3H). LCMS1 3.83min. (M+H)=521. [α]=−126.6° (589 nm, EtOH)

Alternative synthetic schemes for Compound A are included herein aswell.

Example 2 1. Preparation of Compound 1

4-Bromobenzoyl chloride (106.0 kg, 482.9 mol) was dissolved in THF (479L) and cooled to −5˜0° C. Potassium tert-butoxide (75.8 kg, 675.5 mol)was dissolved in THF (565 L) to give a hazy solution, which was cooledto −5° C. and added over 5 h to the acid chloride solution via an inlinefilter so that the temperature was maintained below 5° C.

After 30 minutes, the reaction was assayed for completion by HPLC (<2%starting material). In a separate vessel, NaCl (40 kg) was dissolved inwater (736 L), then heptane (886 L) charged and the mixture was cooledto −5° C. The reaction mixture was added over 5 h to the aqueous mixturemaintaining below 5° C. The reaction vessel was rinsed with heptane (88L) and combined with the batch. After layer separation, the aqueous wasback-extracted with heptane (294 L). The combined organic layer waswashed twice with 426 L water until pH 7, and dried over anhydrous MgSO₄(15.9 kg). The filtrate was concentrated to ˜230 L in vacuum at 30-40°C. Charged THF (609 kg) and concentrated to ˜230 L. This was repeateduntil water<0.05% water and <12% heptane.

¹H NMR (400 MHz, CDCl₃) δ 7.85 (d, J=8.7 Hz, 2H), 7.55 (d, J=8.7 Hz,2H), 1.60 (s, 3H).

¹³C NMR (100 MHz, CDCl₃) δ 165.2, 131.7, 131.2, 131.1, 127.6, 81.7,28.4.

Anal. Calcd for C₁₁H₁₃BrO₂: C, 51.38; H, 5.10; Br, 31.08. Found: C,51.61; H, 5.09; Br, 31.35.

2. Preparation of Compound 3

Sodium tert-butoxide (53.0 kg) in THF (404.6 kg) was de-gassed via threeN₂/vacuum purge cycles and agitated for 15 minutes until the solid wasdissolved at 15-20° C. Palladium acetate (454 g), and DPEphos (1081 g)were then charged under nitrogen. The batch was de-gassed again viathree N₂/vacuum purge cycles and aged for 10 minutes.

4-Chlorovalerophenone (2) (79.0 kg; flushed with THF to remove residualMeOH and H₂O until <0.35% water), tert-butyl bromobenzoate 1 (160.7 kg),and THF (66.0 kg) were then added, and the batch was de-gassed again viathree N₂/vacuum purge cycles. The batch was then heated to 58-64° C. for8 hours and then checked by HPLC for completion (<2% 3).

After cooling to 15-25° C., the batch was quenched into a 0-5° C.mixture of heptane (730.7 kg) and sodium bicarbonate solution (preparedby dissolving 42.8 kg sodium bicarbonate in 808 L water), keeping at0-10° C. The reaction vessel was rinsed with heptane (40.5 kg) andcombined with the mixture. The mixture was warmed to 15-25° C. and thephases separated.

The aqueous phase was back extracted with heptane (385.0 kg). Thecombined organics were poured into a pH 8.5 aqueous solution made upwith 2-mercaptobenzoic acid (32.0 kg), water (354 kg) and stirred at25-30° C. for 6-8 h. After layer separation, the organic phase waswashed twice with 3% Na₂CO₃ solution (549 kg each time). Analysis of theorganic layer indicated that the residual 2-mercaptobenzoic was <0.05%.The organic was washed twice with water (426 kg each time) until pH 7,and further washed twice with 20% NaCl solution (476 kg each time).

A silica plug was prepared in the filter using 50 kg silica 60 wet withcold heptane and topped with anhydrous Na₂SO₄. The organic batch wasthen filtered through the silica gel, and washed with heptane (79.1 kg).

The combined filtrates were concentrated to 160 L yellow oil undervacuum at batch temperature<40° C. 2-Propanol (930 kg) was added and themixture was concentrated to 160 L. Added 2-propanol (620.9 kg) andconcentrated to 160 L at <40° C. The oil was diluted with 2-propanol(231 kg), and warmed to 45-60° C. After stirring for 15 min, H₂O (93 kg)was slowly added at 40-60° C. to the slurry, then allowed it to slowlycool to 22° C. The slurry was then cooled to −5-5° C., aged for 2 h,filtered and washed with 50 kg 2:1 2-propanol/water.

The wet cake was dried under vacuum at 38-40° C./22 in. Hg/N₂ for 22 hto yield product 2 as an off-white solid.

¹HNMR (400 MHz, CDCl₃) δ 7.92 (m, 2H), 7.87 (m, 2H), 7.36 (m, 2H), 7.33(m, 2H), 4.54 (t, J=7.2 Hz, 1H), 2.16 (m, 1H), 1.83 (m, 1H), 1.57 (s,9H), 1.37-1.17 (m, 2H), 0.92 (t, J=7.3 Hz, 3H).

¹³C NMR (100 MHz, CDCl₃) δ 198.4, 165.6, 144.3, 139.6, 135.2, 131.2,130.3, 130.2, 129.1, 128.3, 81.2, 53.7, 36.0, 28.4, 20.9, 14.2.

Anal. Calcd for C₂₂H₂₅ClO₃: C, 70.86; H, 6.76; Cl, 9.51. Found: C,70.73; H, 6.98; Cl, 9.21.

HPLC conditions: Zorbax Eclipse XDB-C8, 4.6×150 mm; A: 0.1% H₃PO₄aqueous; B: acetonitrile; 70% to 95% B over 15 min, hold 2 min, posttime 4 min. 1.0 mL/min, 10 μL, 210 nm, 30° C. column temperature;p-chlorovalerophenone, RT=4.36 min; tert-butyl 4-bromobenzoate, RT=5.56min; product, RT=9.74 min; product acid, RT=3.26 min.

Note: The catalyst used above can be replaced with BINAP or tol-BINAP.

3. Preparation of Compound 4

Ketone 3 (110 kg) in IPA (682 kg) was charged to the hydrogenationvessel, and the solution was thoroughly de-gassed using N₂/vacuum purgecycles.

The catalyst solution was prepared in a separate vessel: potassiumtert-butoxide (7.0 kg) was dissolved in IPA (66 kg) and thoroughlypurged with N₂. The catalyst Ru[(S)-XYL-SEGPHOS][(S)-DIAPEN] (551 g) wasadded, the catalyst mixture aged for 1 hour at 25-30° C., whilst purgingwith N₂. This catalyst preparation was then added to the ketone IPAsolution, taking care to exclude air during this operation, andde-gassing using N₂/vacuum purge cycles after the addition. The batchwas then hydrogenated for 3-5 hours at 20-25° C. using a H₂ pressure of0.64-0.68 Mpa.

The reaction solution was passed through silica gel column (46.4 kg) twotimes. The filtrate was concentrated to ˜880 L by distillation at <40°C. The solution was then heated to 55-58° C., and slowly added water(780 kg) over 1.5 h at the same temperatures. After stirring for 1-1.5h, the mixture was cooled to 20-25° C. over 2-3 h. The slurry was agedfor 2-3 h, then cooled to 0-5° C. over 2-3 h. After stirring for 1-2 h,the slurry was filtered, and the cake was washed twice with cold 2:1IPA:water (230 kg).

The wet cake was dried in 40-45° C. vacuum for 28-30 h, to give theproduct 4 as an off-white solid.

¹H NMR (400 MHz, CDCl₃) δ 7.96 (m, 2H), 7.32 (m, 2H), 7.26 (m, 2H), 7.22(m, 2H), 4.76 (dd, J=7.7, 2.9 Hz, 1H), 2.89 (ddd, J=11.5, 7.7, 4.2 Hz,1H), 1.84 (d, J=2.9 Hz, —OH), 1.62 (s, 9H), 1.61 (m, 1H), 1.41 (m, 1H),1.05 (m, 2H), 0.76 (t, J=7.3 Hz, 3H).

¹³C NMR (100 MHz, CDCl₃) δ 165.9, 146.3, 141.4, 133.7, 131.0, 129.8,128.9, 128.7, 128.4, 81.1, 78.01, 54.2, 34.2, 28.4, 20.6, 14.1.

Anal. Calcd for C₂₂H₂₇ClO₃: C, 70.48; H, 7.26; Cl, 9.46. Found, C,70.45; H, 7.40; Cl, 9.24.

Zorbax Eclipse XDB-C8, 4.6×150 mm; A: 0.1% H₃PO₄ aqueous; B:acetonitrile; 70% to 95% B over 15 min, hold 2 min, post time 4 min. 1.0mL/min, 10 μL, 210 nm, 30° C. column temperature; Major diastereomer,RT=7.74 min; minor diastereomer, RT=6.89 min; Major diastereomer acid,RT=2.66 min; minor diastereomer acid, RT=2.27 min.

Chiral SFC Method: Chiralpak AD-H (250×4 6 mm), isocratic 15% MeOH/CO₂,1.5 mL/min, 200 bar, 35° C., 215 nm, 15 min: desired alcohol, RT=9.8min; enantiomeric alcohol, RT=10.6 min.; diastereomeric alcohols: 5.2 &6.3 min.

4. Preparation of Compound 5

Alcohol 4 (90 kg) was slurried in acetonitrile (600 kg) andorthophosphoric acid (85 wt %, 113.6 kg) charged followed by 60 kgacetonitrile for chasing orthophosphoric acid. The slurry was inertedwith N₂ and heated to 62-68° C., monitoring hourly for conversion.

After 3.5-4 h, reaction was complete by HPLC. The solution was cooled to55-65° C. and water (98 kg) charged over 45 minutes to effectcrystallization. The mixture was cooled to 45-50° C., and seeded with430 g compound 5 to effect crystallization if crystallization has notoccurred. Once a seedbed was established at 45-50° C., further water(861 kg) was charged, and stirred for 1-2 h at these temperatures. Theslurry was cooled slowly to 20-25° C., and then aged for 2-3 h. Theproduct was filtered and washed with 88 kg of 3:1 H₂O/CH₃CN.

The wet cake was dried under vacuum at 38-40° C. for 10-16 h to affordproduct 5 as a yellow solid as a mono-hydrate.

¹H NMR (400 MHz, DMSO-d₆) δ 12.71 (br s, —CO₂H), 7.79 (d, J=8.3 Hz, 2H),7.29 (d, J=8.4 Hz, 2H), 7.19-7.25 (m, 4H), 5.32 (br s, —OH), 4.76 (d,J=6.3 Hz, 1H), 2.85 (dt, J=10.7, 5.4 Hz, 1H), 1.61 (m, 1H), 1.44 (m,1H), 1.00 (m, 2H), 0.73 (t, J=7.3 Hz, 3H).

¹³C NMR (100 MHz, DMSO-d₆) δ 167.4, 147.7, 143.7, 131.0, 129.1, 128.6,128.4, 128.3, 127.6, 75.1, 52.7, 34.0, 20.0, 13.8.

Anal. Calcd for C₁₈H₁₉ClO₃.H₂O: C, 64.19; H, 6.28; Cl, 10.53. Found: C,64.43; H, 6.06; Cl, 10.30.

Zorbax Eclipse Plus C18, 4.6×50 mm; A: 0.1% H₃PO₄ aqueous; B:acetonitrile; 75% to 95% B over 4 min, hold 1 min, post time 2 min. 1.5mL/min, 3 μL, 210 nm, 22° C. column temperature; starting material,RT=2.49 min; product, RT=0.65 min.

Chiral HPLC Method: ChiralPak IB (250×4.6 mm), 0.1% TFA in heptane,B=0.1% TFA in 50/50 EtOH/MeOH, isocratic 6% B, 1.0 mL/min, 10 μL of 1.5mg/mL EtOH, 25° C., 254 nm, 30 min: major enantiomer, RT=17.10 min;minor enantiomer, RT=21.22 min.

5. 2-Bromo-6-fluoro-4-methylaniline 7

6-fluoro-4-methylaniline 6 (15 kg) was added to a mixture of MTBE (92 L,66.6 kg) calcium carbonate (12 kg) and water (135 kg), inerted andcooled to <5° C. Bromine (18.22 kg) was charged, keeping T<10° C. Agefor 30 minutes at T<10° C. The batch was warmed to ˜18° C. and allowedto settle.

The lower aqueous layer was cut away and the organics washed with 5% aqsodium metabisulfite (75 L) then water (75 L). The organic phase wassolvent-switched to toluene (at <40° C. using vacuum), final volume45-60 L, then passed through a silica pad (15 kg) in an oyster filter,washing with toluene (60 L, 52 kg). The filtrates were combined andreduced by vacuum distillation (T<40° C.).

6. 7-Fluoro-5-methyl-1H-indole 8

Method 1:

Palladium allyl chloride dimer (0.74 kg) and DPE Phos (2.17 kg) wereslurried in heptane (165 L, 113 kg) and de-gassed using N₂/vacuumcycles. Dicyclohexylamine (7.85 kg) was added, de-gassed again usingN₂/vacuum cycles, and the resulting suspension was aged for no more than40 minutes.

In another vessel, the bromoaniline 7 (16.5 kg as a 20 wt % toluenesolution), dicyclohexylmethylamine (36.2 kg) and trimethylsilylacetylene (15.1 kg) were de-gassed using N₂/vacuum cycles. The catalystsuspension was then transferred into this vessel over 5 minutes, and thevessel was pressurised to 2300 mbar and heated to 70° C. overnight.

After 17 hours, HPLC analysis showed no starting material present. Thebatch was cooled to 20° C. and filtered through ˜3 kg Solka-Floc in anoyster filter. Heptane (30 L) was used to rinse the vessel then wash thefilter cake. The filtrate and wash were combined and washed with HCl(made up with 15.4 kg conc. HCl and 75 kg water), then water (80 kg).

50 kg silica was loaded into the large oyster filter and wetted withcold heptane (100 L). The organics were then filtered through thesilica, washing/eluting with toluene/heptane (21 kg/38 kg) then toluene(55 kg). The filtrate fractions were combined and distilled to a volumeof 90 L under reduced pressure with maximum batch temperature of 50° C.

Tetrabutylammonium fluoride trihydrate (27.8 kg) was dissolved in THF(79 kg) and methanol (4.1 L), and the indole toluene solution added toit over 5 minutes. The solution was heated to 80-85° C. and THF removedby atmospheric distillation.

Once reaction was complete, distillation was continued at 100-150 mbarto remove residual THF. The batch was then cooled to 20° C. and washedwith 5 wt % brine (44 kg). The brine wash was extracted with toluene (42kg). The combined toluene organics were then washed with 5 wt % brine.The organics were filtered and the filtrate concentrated to give atoluene solution of indole 8.

¹H NMR (400 MHz, CDCl₃) δ 8.20 (br s, —NH), 7.22 (d, J=0.6 Hz, 1H), 7.20(dd, J=2.9, 2.7 Hz, 1H), 6.78 (dd, J=12.0, 0.7 Hz, 1H), 6.52 (ddd,J=3.3, 3.3, 2.2 Hz, 1H), 2.46 (s, 3H).

¹³C NMR (100 MHz, CDCl₃) δ 149.4 (d, J=242.9 Hz), 131.8 (d, J=5.5 Hz),130.2 (d, J=5.7 Hz), 125.0, 122.6 (d, J=13.2 Hz), 116.1 (d, J=3.1 Hz),108.6 (d, J=15.7 Hz), 103.0 (d, J=2.3 Hz), 21.6.

¹⁹F NMR (376 MHz, CDCl₃) δ −136.5.

Anal. Calcd for C₉H₈FN: C, 72.47; H, 5.41; F, 12.74; N, 9.39. Found: C,72.15; H, 5.34; F, 12.74; N, 9.28.

Zorbax Eclipse XDB-C8, 4.6×150 mm; A: 0.1% H₃PO₄ aqueous; B:acetonitrile; 70% to 95% B over 15 min, hold 2 min, post time 4 min. 1.0mL/min, 10 μL, 210 nm, 30° C. column temperature; product, RT=2.67 min.

Method 2:

Step 1: Sugasawa Reaction

Reagent MW Amount mmol Equiv. 2-Fluoro-4-methylaniline 125.15 12.52 g100 2.00 Chloroacetonitrile 75.5 3.78 g  50 1.00 BCl₃/CH₂Cl₂ (1.0M) 110mL 110 2.20 AlCl₃ pellet 133.34 8.00 g  60 1.20 1,2-Dichloroethane (DCE)125 mL 10 V of aniline HCl (2.0M in water) 80 mL 160Product-chloroacetophenone 201.63 10.08 g  50 100%

A vessel was cooled via ice bath, was charged with AlCl₃ pellets, 100 mLDCE, and BCl₃/DCM solution 110 mL under N₂ with an outlet into anaqueous NaOH solution bath. A solution of SM-aniline was then addeddropwise with caution to keep t<20 C. Upon completion, the grey slurrywas added SM-nitrile in one portion, and extra DCE was used to rinse theRBF. The ice bath was removed, and the reaction was heated to reflux.

After 6 h of reflux, the mixture was cooled, quenched with 2N HCl 80 mL,and heated to reflux for 20 min. The mixture was extracted with DCM,washed with 2N HCl, water, and brine. After concentration, the solid wasstirred with 25 mL hexanes, cooled via an ice bath, filtered, and washedwith extra 50 mL hexanes, A dark yellow solid was obtained.

The combined HCl wash was treated with NaOH (pellets, 14.7 g) to pH=5-6,extracted with MTBE/Heptane (1:1) and concentrated.

Step 2: Reduction and Cyclization

Reagent MW Amount mmol Equiv. chloroacetophenone (96.6%) 201.63 2033 mg9.74 1.00 NaBH₄  37.83 456 mg 12.0  2.46 1,4-Dioxane 20 mL 10 V Water2.0 mL  1 V Product-indole 149.16 1453 mg 9.74 100%

Chloroacetophenone was dissolved in dioxane followed by addition ofNaBH₄ and water. The mixture was aged at room temperature for 25 minbefore heated to reflux for 2 h, cooled to room temperature, worked upwith MTBE and water. Concentration of the workup gave a brown clearliquid, which solidified upon freezing.

The formed indole was used to prepare nosyl indole as a white solidpowder after recrystallization from MeCN/water (2:1).

7. 7-Fluoro-5-methyl-1-[(4-nitrophenyl)sulfonyl]-1H-indole 8

Indole 8 (3.2 kg as a 10 wt % solution in toluene) was distilled to avolume of 90 L under reduced pressure with a maximum batch temperatureof 50° C. 48 wt % sodium hydroxide (15 kg, 9.6 L) was charged. The batchwas cooled to 15° C. and tetrabutylammonium sulphate (363 g) added.Nosyl chloride (5.92 kg) was dissolved in toluene (15.6 kg) and thesolution charged slowly, keeping T<25° C. After 10 minutes, HPLCanalysis showed complete reaction.

Cold (10° C.) 5 wt % sodium hydrogen carbonate solution (45 L) wascharged and the phases separated. The organics were washed with 5 wt %sodium hydrogen carbonate solution (31 L), and then washed with HCl (30kg water & 730 mL conc HCl). The organics were distilled to a volume of˜45 L under reduced pressure with maximum batch temperature of 40° C.Isopropanol (35 kg, 45 L) was added, and the solution passed through theCUNO carbon filtration system, using a 3.5 kg R-55SLP cartridge.

Isopropanol (19.3 kg, 25 L) was added to the filtrate and it wasdistilled under reduced pressure with maximum batch temperature of 40°C., adding additional isopropanol until <10 mol % toluene is present byNMR, to a final volume of ˜10 wt % product in IPA. The batch was cooledto 5° C. to crystallise the product. The slurry was filtered, and washedwith isopropanol (30 L, 24 kg), then cold IPA/water (24 kg/1.5 kg). Thesolid was dried on the filter under a nitrogen stream to give theproduct 9 as a brown solid.

¹H NMR (400 MHz, CDCl₃) δ 8.32 (d, J=8.8 Hz, 1H), 8.11 (d, J=8.8 Hz,1H), 7.70 (d, J=3.7 Hz, 1H), 7.13, (s, 1H), 6.81 (d, J=13.1 Hz, 1H),6.66 (dd, J=3.5, 2.4 Hz, 1H), 2.38 (s, 3H);).

¹³C NMR (100 MHz, CDCl₃) δ 150.8, 149.2 (d, J=249.2 Hz), 144.1, 135.5(d, J=6.5 Hz), 135.3 (d, J=3.8 Hz), 129.2 (d, J=2.8 Hz), 128.7, 128.6,120.0 (d, J=11.2 Hz), 117.6 (J=3.4 Hz), 113.1 (d, J=19.4 Hz), 109.1 (d,J=2.1 Hz).

Anal. Calcd for C₁₅H₁₁FN₂O₄S: C, 53.89; H, 3.32; F, 5.68; N, 8.38; 0,19.14; S, 9.59. Found: C, 53.68; H, 3.16; F, 5.58; N, 8.30; S, 9.64.

Zorbax Eclipse XDB-C8, 4.6×150 mm; A: 0.1% H₃PO₄ aqueous; B:acetonitrile; 70% to 95% B over 15 min, hold 2 min, post time 4 min. 1.0mL/min, 10 μL, 210 nm, 30° C. column temperature; starting indole,RT=2.49 min; N-(4-nosyl)-indole, RT=3.74 min.

8. 4-[(1S)-1-((R)-(4-Chlorophenyl){7-fluoro-5-methyl-1-[(4-nitrophenyl)sulfonyl]-1H-indol-3-yl}methyl)butyl]benzoicacid 10

Method 1:

Acid 5 (8.2 kg of 98 wt %) and nosyl indole 9 (8.86 kg, 96 wt %) wereslurried in dichloromethane (160 kg), inerted and cooled to 15° C.BF₃OEt₂ (10.74 kg) was charged, keeping T<25° C. The brown solution wasaged at 20° C. for 18 h, then cooled to 15° C. and water (80 kg) added.

The biphasic mixture was filtered through a pad of Solka-Floc (˜5 kg)contained in an oyster filter, and the pad was washed with 10 kgdichloromethane. The filtrate was separated and the lower DCM organicwashed with water (80 kg).

The DCM organics were then stirred gently overnight with MP-TMT resin(2.4 kg), then filtered, washing with IPAC (30 L).

The filtrate volume was then reduced by vacuum distillation (T<35° C.).Isopropyl acetate (105 kg) was added, and this solution was filtered,washing with IPAC and DCM.

The IPAC solution was warmed to 50° C. and heptane (38 kg) charged,keeping T at approximately 45° C. Seed (10 g) was charged, and the batchcooled to 20° C. overnight (using a linear ramp on the PCS system).Further seeding or heptane addition caused crystallisation.

The product was dried in vacuo to give 9 as a tan solid.

Method 2:

To a mixture of alcohol 5 (42.7 kg, 93.7 wt %) and indole 9 (41.3 kg,99.5 wt %) was added trifluoroacetic acid (943 kg) and methanesulfonicacid (6.0 kg). After stirring for 19 h at 22° C., the solution wasdiluted with isopropyl acetate (1070 kg) (temperature controlled from10-25° C.), cold 20 wt % NaOH (1200 kg), and 15 wt % K₂HPO₄ (1430 kg).After separating the layers, the organic was pH adjusted to a pH of 2with 0.1 N HCl (136 kg). After separating the layers, the organic waswashed with water (1207 kg).

The organic phase was filtered and then washed with IPAC. The organicphase was concentrated and flushed with IPAC to remove water (target:<200 ug/mL).

The batch was seeded with 0.5 wt % (relative to products) at 35° C. andaged for 60 min, then cooled 20° C. over 1 h to allow a seed bed todevelop. Heptane was added over 300 min. The slurry was filtered, washedwith 1:4 IPAC/heptane (430 kg), and vacuum dried at 40° C. overnight toafford product 10 as a tan solid.

4-[(1S)-1-((R)-(4-chlorophenyl){7-fluoro-5-methyl-1-[(4-nitrophenyl)sulfonyl]-1H-indol-3-yl}methyl)butyl]benzoicacid-isopropyl acetate (1:1)

Major Diastereomer:

¹H NMR (400 MHz, CDCl₃) δ 8.13 (d, J=8.9 Hz, 2H), 8.05 (d, J=8.3 Hz,2H), 7.52 (d, J=8.7 Hz, 2H), 7.45 (s, 1H), 7.40 (d, J=8.3 Hz, 2H),7.38-7.31 (m, 4H), 6.89 (s, 1H), 6.68 (d, J=12.8 Hz, 1H), 4.32 (d,J=11.5 Hz, 1H), 3.43 (dt, J=10.8, 3.5 Hz, 1H), 3.29 (s, 3H), 1.55 (m,1H), 1.47 (m, 1H), 1.06 (m, 2H), 0.76 (t, J=7.3 Hz, 3H).

¹³C NMR (100 MHz, CDCl₃) δ 171.4, 151.0, 150.7, 149.1 (d, J=250.6 Hz),143.4, 140.1, 135.4 (d, J=6.4 Hz), 134.9 (d, J=3.2 Hz), 133.1, 130.7,129.9, 129.3, 128.6, 128.5, 127.7, 127.7, 126.4 (d, J=3.2 Hz), 124.5,124.4, 119.8 (d, J=11.1 Hz), 115.5, 113.5 (d, J=19.3 Hz), 50.5, 47.7,37.4, 21.4, 20.4, 14.1.

¹H NMR (400 MHz, DMSO-d₆) δ 8.23 (d, J=8.8 Hz, 2H), 7.95 (s, 1H), 7.83(d, J=8.2 Hz, 2H), 7.70-7.60 (m, 4H), 7.57 (d, J=8.2 Hz, 2H), 7.39 (d,J=8.4 Hz, 2H), 7.27 (s, 1H), 6.83 (d, J=13.2 Hz, 1H), 4.61 (d, J=11.8Hz, 1H), 3.80 (dt, J=11.6, 3.2 Hz, 1H), 2.25 (s, 3H), 1.45 (m, 1H), 1.29(m, 1H), 0.93 (m, 2H), 0.68 (t, J=7.3 Hz, 3H).

¹³C NMR (100 MHz, DMSO-d₆) δ 171.8, 167.2, 150.4, 149.9, 148.2 (d,J=250.0 Hz), 141.7 (d, J=30.3 Hz), 135.0 (d, J=6.5 Hz), 134.8 (d, J=3.2Hz), 131.1, 130.2, 129.1, 128.5, 128.4, 128.3, 128.1, 126.8, 125.5 (d,J=1.6 Hz), 124.7, 118.5 (d, J=10.3 Hz), 115.7, 112.7 (d, J=19.5 Hz),47.9, 45.8, 36.7, 20.6, 19.7, 13.6.

¹⁹F NMR (376 MHz, CDCl₃) δ −122.8.

Anal. Calcd for C₃₈H₃₈C1FN₂O₈S: C, 61.91; H, 5.20; Cl, 4.81; F, 2.58; N,3.80; S, 4.35. Found: C, 62.04; H, 5.09; N, 3.79; Cl, 4.82; F, 2.63; S,4.47.

Minor Diastereomer:

Selective NMR signals: ¹H NMR (400 MHz, CDCl₃) δ 8.32 (d, J=8.9 Hz, 2H),8.08 (d, J=8.9 Hz, 2H), 7.94 (d, J=8.2 Hz, 2H), 7.72 (s, 1H), 7.16 (d,J=8.2 Hz, 2H), 7.07 (s, 1H), 7.05 (d, J=8.5 Hz, 2H), 6.98 (d, J=8.5 Hz,2H), 6.68 (d, J=12.8 Hz, 1H), 4.27 (d, J=10.8 Hz, 1H), 3.47 (m, 1H),2.37 (s, 3H), 1.85 (m, 1H), 1.20 (m, 1H), 1.15 (m, 2H), 0.83 (t, J=7.3Hz, 3H); ¹⁹F NMR (376 MHz, CDCl₃) δ −122.3; Anal. Calcd forC₃₃H₂₈C1FN₂O₆S.0.5 MTBE.0.2 heptane: C, 63.38; H, 5.36; N, 4.01; Cl,5.07; F, 2.72; S, 4.59. Found: C, 63.59; H, 5.36; N, 3.84; Cl, 4.94; F,2.75; S, 4.61.

Zorbax Eclipse XDB-C8, 4.6×150 mm; A: 0.1% H₃PO₄ aqueous; B:acetonitrile; 70% to 95% B over 15 min, hold 2 min, post time 4 min. 1.0mL/min, 10 μL, 210 nm, 30° C. column temperature; Major diastereomer,RT=7.23 min; minor diastereomer, RT=5.66 min; 3-tert-butylated indole 5,RT=6.69 min.

Chiral SFC Method: ChiralPak IB column (250×4.6 mm), isocratic 25% MeOHw 0.1% TFA/CO2, 1.5 ml/min, 200 bar, 35 C, 35 min; Enantiomer of desired(R,S), RT=17.28; Desired (S,R), RT=18.11 min; Diastereomer of desired(S,S), RT=19.85 min; Enantiomer of diastereomer of desired (R,R),RT=24.15 min.

9. 3-[(4-{(1S)-1-[(R)-(4-chlorophenyl)(7-fluoro-5-methylindol-3-yl)methyl]butyl}benzoyl)amino]-propanoic acid 12

Penultimate compound 10 (7.40 kg) was dissolved in THF (32.9 kg) anddegassed using N₂/vacuum purge cycles. N,N-carbonyldiimidazole (2.64 kg)was charged, and the batch was warmed to 40° C. for 45 minutes. HPLCshowed complete conversion to acylimidazolide.

The batch was cooled to 30° C. and β-alanine methyl ester (2.60 kg)added. The batch was then heated to 60° C. for 4 hours. HPLC analysisshowed complete conversion to amide/ester.

2.5 M NaOH (32.6 L; prepared from 5.70 kg 10 M NaOH and 24.5 kg water)was charged and the batch aged at 60° C. for 1.5 h, then further 10 MNaOH (1.63 kg) added and the batch aged at 60° C. overnight. The batchwas cooled to 25° C. and the lower aqueous layer cut away. Further water(14.1 kg) and 10 M NaOH (2.78 kg) were charged, and the batch heated to60° C. for 2 hours.

The batch was cooled to 25° C. and MTBE (48.8 L, 36.1 kg) charged, then5 wt % NaCl (2.44 kg NaCl in 48.8 kg water). After agitation andsettling, the phases were separated and the organics washed with 5 wt %NaCl (2.44 kg NaCl in 48.8 kg water), 3 M HCl (2.68 kg conc. HCl in 9.8kg water) and water (48.8 kg).

The organics were concentrated to a volume of 15 L using vacuum with amaximum batch temperature of 30° C. Isopropanol (47.4 L, 37.2 kg) wasthen added and the batch concentrated to 15 L again. Isopropanol (27 L,21 kg) was added and the solution was filtered, washing withisopropanol. The filtrate was then vacuum distilled with maximum batchtemperature of 30° C. Water (26.6 kg) was added, seed added and theslurry aged for 45 minutes.

The slurry was filtered, washing with IPA/water (4.6 kg/11.8 kg) thenwater (35 kg). The solid was dried.

The cake was transferred to a vacuum oven at 42° C. for 24 h. The driedsolids were passed through a co-mill to give product 12 (98.7 LCAP, 101LCWP) as an off-white solid.

Amorphous Material

The amorphous phase of compound A was obtained by lyophilization. It canalso be obtained by other techniques such as spraying drying and meltquenching. Amorphous material can be detected in samples as shown inFIG. 16, as part of a mixture of amorphous and polymorphic materials.

Polymorphic Materials

The polymorphic forms of compound A were prepared using material thatwas produced as described in steps 1 through 9. Seed crystal wasobtained from lab scale preparations spontaneously or uponchromatographic purification. An X Ray Powder Diffraction Pattern of amixture of amorphous and polymorphic forms is included for comparisonpurposes as FIG. 16.

X-ray powder diffraction (XRPD) patterns for the solid phases ofcompound A were generated on a Philips Analytical X'Pert PRO X-rayDiffraction System with PW3040/60 console. The diffraction peakpositions were referenced by silicon which has a 2 theta value of 28.443degree. A PW3373/00 ceramic Cu LEF X-ray tube K-Alpha radiation was usedas the source. The experiments were run at ambient condition except forthe anhydrate of polymorphic form I, where the measurement was carriedout at 5% relative humidity at room temperature.

Solid-state carbon-13 NMR spectra were obtained on a Bruker DSX 400WBNMR system using a Bruker 4 mm double resonance CPMAS probe. Thecarbon-13 NMR spectrum utilized proton/carbon-13 cross-polarizationmagic-angle spinning with variable-amplitude cross polarization. Thesample was spun at 8 kHz. Chemical shifts are reported on the TMS scaleusing the carbonyl carbon of glycine (176.03 p.p.m.) as a secondaryreference.

Solid-state fluorine-19 NMR spectra were obtained on a Bruker DSX 500WBNMR system using a Bruker 4 mm CRAMPS probe. The NMR spectrum utilizedproton/fluorine-19 cross-polarization magic-angle spinning withvariable-amplitude cross polarization. The samples were spun at 15.0kHz. A vespel endcap was utilized to minimize fluorine background.Chemical shifts are reported using poly(tetrafluoroethylene) (Teflon) asan external secondary reference which was assigned a chemical shift of−122 ppm.

Differential Scanning Calorimetry

TA Instruments DSC 2910 or equivalent instrumentation is used to conductdifferential scanning calorimetry. Between 1 and 5 mg of a sample isweighed into a sample pan and placed at the sample position in thecalorimeter cell. An empty pan is placed at the reference position(either the sample and reference pans are closed or both are open). Thecalorimeter cell is closed and a flow of nitrogen is passed through thecell. The heating program is set to heat the sample at a heating rate of10° C./min to a temperature of ˜300° C. The heating program is started.At the completion of the run, the data are analyzed using the Universalanalysis program contained in the system software. Integration of anyendotherms present is carried out between points on the baseline attemperatures above and below the range in which the endothermic event(s)are observed. Reported data include observed onset temperature, peaktemperature, and associated enthalpy of melting.

Thermogravimetric Analysis (TG):

An instrument such as Perkin Elmer model TGA 7 or equivalent is used toconduct thermogravimetric analysis. Experiments are performed under aflow of nitrogen at a heating rate of 10° C./min, and samples are heatedto a temperature of about 300° C. To start an experiment, the balance istared, approximately 3 to 20 mg of sample is added to the platinum pan,the furnace is raised, the sample is weighed, and the heating program isstarted. Weight/temperature data are collected automatically by theinstrument and can be converted to weight percent/temperature. Analysisof the results is carried out by selecting the Delta Y function withinthe instrument software and the weight loss up to the completion ofmelting is reported.

Hydrate

The hydrate was produced in accordance with the process set forth above.The term hydrate refers to different crystalline forms of the compound,such as the monohydrate, dihydrate, hemi-hydrate, and the like.

Hydrate: ¹H NMR (400 MHz, CDCl₃) δ 8.20 (br s, NH), 7.46 (d, J=8.1 Hz,2H), 7.28 (d, J=8.5 Hz, 2H), 7.25 (d, J=8.5 Hz, 2H), 7.17 (d, J=8.1 Hz,2H), 6.94 (s, 1H), 6.83 (d, J=2.1 Hz, 1H), 6.72 (t, J=6.0 Hz, —CONH),6.60 (d, J=11.9 Hz, 1H), 4.37 (d, J=10.7 Hz, 1H), 3.58 (m, 2H), 3.37(dt, J=10.7, 3.2 Hz, 1H), 2.57 (m, 2H), 2.35 (s, 3H), 1.50 (m, 1H), 1.40(m, 1H), 0.98 (m, 2H), 0.71 (t, J=7.3 Hz, 3H).

¹³C NMR (100 MHz, CDCl₃) δ 176.6, 168.2, 149.2 (d, J_(CF)=243.8 Hz),149.0, 142.6, 132.1, 131.7, 130.9 (d, J_(CF)=5.5 Hz), 130.0, 129.6 (d,J_(CF)=5.6 Hz), 128.8, 128.5, 127.1, 122.8, 122.6 (d, J_(CF)=13.3 Hz),118.6 (d, J_(CF)=1.2 Hz), 114.3 (d, J_(CF)=2.5 Hz), 108.6 (d,J_(CF)=15.7 Hz), 50.3, 48.2, 37.3, 35.5, 34.0, 21.8, 20.5, 14.1.

¹⁹F NMR (376 MHz, CDCl₃) δ −136.3.

Anal. Calcd for C₃₀H₃₁ClFN₂O₃₅: C, 67.98; H, 5.90; Cl, 6.69; F, 3.58; N,5.29. Found: C, 68.11; H, 5.84; Cl, 6.70; F, 3.54; N, 5.33.

Zorbax Eclipse XDB-C8, 4.6×150 mm; A: 0.1% H₃PO₄ aqueous; B:acetonitrile; 70% to 95% B over 15 min, hold 2 min, post time 4 min. 1.0mL/min, 10 μL, 210 nm, 30° C. column temperature; desired, RT=3.65 min.

Chiral SFC LC: Chiralpak AD-H column (250×4 6 mm), isocratic 15% MeOHwith 25 mM isobutylamine/CO2, 1.5 mL/min, 200 bar, 35° C., 210 nm, 30min run time: Desired, RT=18.9 min; enantiomer of desired, RT=15.7 min.

Hydrate/Methanolate

One aspect of the invention that is of interest relates to a crystallinepolymorphic compound of formula A in the form of a free acidhydrate/methanolate solvate.

The hydrate/methanolate solvate of compound A was obtained by directcrystallization of compound A in methanol/water at different solventcompositions.

Alternatively, it can be obtained by adding excess amount of compound Ain methanol/water, stirring for several hours, and recovering the solidsby filtration. Characterization is in accordance with Table 1 below.

TABLE 1 X-ray powder diffraction: free acid hydrate/ methanolate ofCompound A 2θ(2 theta)(degrees) d-spacing (Å)  7.5 11.74   8.8 10.10 11.3 7.84 13.7 6.44 14.9 5.95 15.4 5.74 17.6 5.05 18.4 4.83 20.3 4.3721.3 4.17 23.4 3.81 25.6 3.48Hydrate

An aspect of the invention that is of interest relates to a crystallinepolymorphic form of a compound of formula A in the form of a free acidhydrate.

The hydrate of compound A was obtained by removing methanol from thehydrate/methanolate. This can be achieved by drying thehydrate/methanolate at 40° C. at >60% relative humidity. The hydrate canalso be converted from the anhydrate of polymorphic form I of compound Aby exposing the anhydrate to an atmosphere of >20% relative humidity atroom temperature. X Ray characterization is as in Table 2 below.

FIG. 1 is the X-ray powder diffraction (XRPD) pattern for the free acidhydrate polymorphic Form I of Compound A; with selected d-spacingslisted in Table 2 below. The compound of formula A exhibits at leastthree of the d-spacings shown in Table 2, and preferably more thanthree.

TABLE 2 XRPD: free acid hydrate polymorphic Form I of Compound A 2θ(2theta)(degrees) d-spacing (Å)  7.5 11.74   8.8 10.10  11.3 7.84 13.76.44 14.9 5.95 15.4 5.74 17.6 5.05 18.4 4.83 20.3 4.37 21.3 4.17 23.43.81 25.6 3.48

FIG. 2 is the solid-state fluorine-19 CPMAS NMR spectrum of the freeacid hydrate polymorphic Form I of Compound A. Form I exhibitedcharacteristic signals with chemical shift values of −132.7 and −134.3ppm.

FIG. 3 is the solid-state carbon-13 CPMAS NMR spectrum of the free acidhydrate polymorphic Form I of Compound A. Form I exhibitedcharacteristic signals with chemical shift values of 11.3, 20.2, 23.9,33.1, 45.5, 108.6, 118.7, 131.7, 139.3, 148.5, 170.1 and 179.9 ppm.

Another aspect of the invention that is of interest relates to thecrystalline free acid hydrate of a compound of formula A wherein atleast three of the X-ray powder diffraction pattern d-spacings are asfound in Table 2.

Another aspect of the invention that is of interest relates to thecrystalline free acid hydrate of a compound of formula A wherein atleast five of the X-ray powder diffraction pattern d-spacings are asfound in Table 2.

Another aspect of the invention that is of interest relates to thecrystalline free acid hydrate of a compound of formula A wherein atleast three C13 solid state NMR characteristic signals with chemicalshift values are selected from the group consisting of: 11.3, 20.2,23.9, 33.1, 45.5, 108.6, 118.7, 131.7, 139.3, 148.5, 170.1 and 179.9ppm.

Anhydrate Form I:

Another aspect of the invention that is of interest relates to acrystalline polymorphic compound of formula A in the form of ananhydrous free acid.

More particularly, an aspect of the invention that is of interestrelates to a crystalline polymorphic compound of formula A in the formof an anhydrous free acid, said polymorphic compound having at leastthree X-ray powder diffraction pattern d spacings in accordance withTable 3.

Even more particularly, an aspect of the invention that is of interestrelates to a crystalline polymorphic compound of formula A in the formof an anhydrous free acid, said polymorphic compound having at leastfive X-ray powder diffraction pattern d spacings in accordance withTable 3.

Another aspect of the invention relates to a crystalline polymorphiccompound of formula A in the form of an anhydrous free acid, saidpolymorphic compound having a 19F solid state NMR as shown in FIG. 5.

The anhydrate I of compound A was obtained either fromhydrate/methanolate of compound A by removing methanol/water or fromhydrate of compound A by removing water. This can be achieved by dryingthe hydrate/methanolate or the hydrate with N₂ at room temperature or athigher temperatures. Anhydrate form I of Compound A exhibits at leastthree of the d-spacings shown in Table 3, and preferably more thanthree. The solid state 19F NMR characterization for anhydrous free acidpolymorphic form I of Compound A exhibited characteristic signals withchemical shift values of −134.8 and −136.3 ppm.

TABLE 3 X-ray powder diffraction: anhydrous free acid polymorphic Form Iof Compound A 2θ(2 theta)(degrees) d-spacing (Å)  7.8 11.35   8.7 10.19  9.7 9.11 11.3 7.82 11.9 7.41 14.9 5.93 15.7 5.63 17.4 5.10 20.1 4.4121.7 4.10 25.6 3.47Anhydrate Form II:

Another aspect of the invention that is of interest relates to acrystalline polymorphic compound in accordance with formula A in theform of a free acid anhydrate of form II.

In particular, an aspect of the invention that is of interest relates toa crystalline polymorphic compound in accordance with formula A in theform of a free acid anhydrate of form II, wherein at least three of thex-ray powder diffraction pattern d-spacing peaks are in accordance withtable 4.

Even more particularly, an aspect of the invention that is of interestrelates to a crystalline polymorphic compound in accordance with formulaA in the form of a free acid anhydrate of form II, wherein at least fiveof the x-ray powder diffraction pattern d-spacing peaks are inaccordance with table 4.

Anhydrate Form II is prepared from a solution of the crystalline hydrateform at a concentration of >100 mg/mL in acetonitrile. The solution issonicated for approximately 5 minutes to induce crystallization.Anhydrate II is metastable. It converts to crystalline Anhydrate IIIwhen heated to 160° C. for 1 hour or in solutions of ethanol, isoamylalcohol and isopropyl acetate. The X Ray Powder Diffraction peaks areshown below in Table 4. Anhydrate form II of compound A exhibits atleast 3 of the peaks shown below in table 4, and preferably more thanfour.

TABLE 4 X-Ray Powder Diffraction peaks for Anhydrate Form II d-spacing[Å] 2θ(2 theta)(degrees) 3.59 24.82 4.05 21.95 4.10 21.70 4.53 19.584.72 18.79 5.93 14.93 7.02 12.62 7.31 12.11 8.07 10.96 12.36   7.15

The thermogravimetric analysis curve for crystalline anhydrate form IIis shown in FIG. 7.

The differential scanning calorimetry (DSC) curve of the crystallineanhydrate Form II of Compound I of the present invention is shown inFIG. 8.

Another aspect of the invention that is of interest relates to acrystalline polymorphic compound of formula A of Form II, having atleast three C13 solid state NMR peaks selected from the group consistingof: 131.51, 130.07, 127.68, 126.30, 123.81, 121.65, 52.57, 39.62, 31.77,and 20.79 ppm,

The 13 CPMAS NMR spectrum for the crystalline anhydrate Form II ofCompound A is shown in FIG. 9. Characteristic peaks for anhydrate FormII are observed at 131.51, 130.07, 127.68, 126.30, 123.81, 121.65,52.57, 39.62, 31.77, and 20.79 ppm.

FIG. 10 shows the fluorine-19 spectra (spinning sideband patterns) foranhydrate Form II of compound I. Center band peaks for anhydrate Form IIare observed at −134.80 and −137.08 ppm.

Anhydrate Form III

Another aspect of the invention that is of interest relates to acrystalline polymorphic compound in accordance with formula A in theform of a free acid anhydrate of form III.

In particular, an aspect of the invention that is of interest relates toa crystalline polymorphic compound in accordance with formula A in theform of a free acid anhydrate of form III, wherein at least three of thex-ray powder diffraction pattern d-spacing peaks are in accordance withtable 5.

Even more particularly, an aspect of the invention that is of interestrelates to a crystalline polymorphic compound in accordance with formulaA in the form of a free acid anhydrate of form III, wherein at leastfive of the x-ray powder diffraction pattern d-spacing peaks are inaccordance with table 5.

Anhydrate Form III is prepared by heating anhydrate form II to 160° C.for 1 h. It can also be prepared by slurrying the hydrate or theAnhydrate II in ethanol, isoamyl alcohol and isopropyl acetate.

X-Ray Powder Diffraction for Anhydrate Form III

FIG. 11 is a characteristic X-ray diffraction pattern of the crystallineanhydrate Form III of Compound I of the present invention.

The anhydrate Form III exhibited characteristic reflectionscorresponding to d-spacings are shown below in Table 5. Anhydrate formIII of compound A exhibits at least three of the d-spacings shown intable 5, and preferably more than three.

TABLE 5 2 theta d-spacing [Å] 26.42 3.37 24.90 3.58 24.61 3.62 23.873.73 23.21 3.83 22.44 3.96 19.66 4.52 19.43 4.57 17.94 4.94  12.29277.20Thermogravimetric Analysis for Anhydrate Form III

FIG. 12 is a typical thermogravimetric analysis curve of the crystallineanhydrate Form III of Compound A.

Differential Scanning Calorimetry

FIG. 13 is a typical differential scanning calorimetry (DSC) curve ofthe crystalline anhydrate Form III of Compound A.

Solid State NMR

Another aspect of the invention that is of interest relates to acrystalline polymorphic compound of formula A of Form III, having atleast three C13 solid state NMR peaks selected from the group consistingof: 177.58, 148.52, 146.17, 130.34, 121.11, 113.44, 52.77, 31.30, 21.42,and 14.04 ppm.

FIG. 14 shows the solid state carbon-13 CPMAS NMR spectrum for thecrystalline anhydrate Form III of Compound A.

Anhydrate III can be characterized by peaks at 177.58, 148.52, 146.17,130.34, 121.11, 113.44, 52.77, 31.30, 21.42, and 14.04 ppm.

FIG. 15 shows the fluorine-19 spectra (spinning sideband patterns) ofcrystalline Anhydrate III of Compound A. Anhydrate form III exhibitscenter band signals at −132.96 and −136.45 ppm.

Another aspect of the invention that is of interest relates to apharmaceutical composition that is comprised of a polymorphic form of acompound of formula A in combination with a pharmaceutically acceptablecarrier.

The following are examples of pharmaceutical dosage forms containing acompound of Formula A:

Injectable Mg/ Suspension (im.) mg/mL Tablet tablet Compound of FormulaA 10.0  Compound of Formula A 25.0 Methylcellulose 5.0 MicrocrystallineCellulose 415 Tween 80 0.5 Povidone 14.0 Benzyl alcohol 9.0Pregelatinized Starch 4.35 Benzalkonium chloride 1.0 Magnesium Stearate2.5 Water for injection t.d. Total 500 mg 1.0 mL mg/ Per Capsule capsuleAerosol Canister Compound of Formula A 25.0 Compound of Formula A 250 mgLactose 735 Lecithin, NF Liq. Conc.  1.2 mg Mg Stearate 1.5Trichloromethane, NF 4.025 g Total 600 mg Dichlorodifluoromethane, 12.15g NF

Certain embodiments of the invention has been described in detail;however, numerous other embodiments are contemplated as falling withinthe invention. Thus, the claims are not limited to the specificembodiments described herein. All patents, patent applications andpublications that are cited herein are hereby incorporated by referencein their entirety.

What is claimed is:
 1. A process for synthesizing a compound of formulaA:

comprising deprotecting a compound of formula 11a:

wherein P¹ and P² represent protecting groups, to produce a compound offormula A.
 2. A process for synthesizing a compound of formula A:

comprising reacting a compound of formula 10a:

wherein P¹ represents a protecting group with a beta alanine ester ofthe formulaH₂NCH₂CH₂CO₂P² wherein P² represents a protecting group, with a peptidecoupling agent, to produce a compound of formula 11a:

and deprotecting compound 11a to produce a compound of formula A.
 3. Aprocess for the synthesis of a compound of formula A:

comprising reacting a compound of the formula 11:

with base to produce a compound of formula A.
 4. A process in accordancewith claim 3 wherein the base is NaOH.